Recombinant Human Lactate Dehydrogenase B/LDHB Protein, CF

R&D Systems | Catalog # 9205-HB

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Key Product Details

  • R&D Systems E. coli-derived Recombinant Human Lactate Dehydrogenase B/LDHB Protein (9205-HB)
  • Quality control testing to verify active proteins with lot specific assays by in-house scientists
  • All R&D Systems proteins are covered with a 100% guarantee

Source

E. coli

Accession Number

P07195

Applications

Enzyme Activity
View all Lactate Dehydrogenase B/LDHB Proteins and Enzymes »

Product Specifications

Source

E. coli-derived human Lactate Dehydrogenase B/LDHB protein
Ala2-Leu334, with N-terminal Met and 6-His tag

Purity

>95%, by SDS-PAGE visualized with Silver Staining and quantitative densitometry by Coomassie® Blue Staining.

Endotoxin Level

<1.0 EU per 1 μg of the protein by the LAL method.

N-terminal Sequence Analysis

Met

Predicted Molecular Mass

37 kDa

SDS-PAGE

36 kDa, reducing conditions

Activity

Measured by its ability to reduce pyruvate to lactate.
The specific activity is >60,000 pmol/min/μg, as measured under the described conditions.

Formulation, Preparation, and Storage

9205-HB
Formulation Supplied as a 0.2 μm filtered solution in Tris, NaCl, TCEP and Glycerol.
Shipping The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.
Stability & Storage Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 6 months from date of receipt, -20 to -70 °C as supplied.
  • 3 months, -20 to -70 °C under sterile conditions after opening.

Background: Lactate Dehydrogenase B/LDHB

A hallmark of most cancer cells is an altered metabolism involving a shift to aerobic glycolysis with lactate production coupled with a higher uptake of glucose as the main source of energy. Lactate dehydrogenase (LDH) is key to this shift by catalyzing the formation of lactate by reducing pyruvate with NADH, which also generates NAD(+) that is essential for the continuity of glycolysis (1). Inhibiting LDH activity has an anti-proliferative effect on cancer cells (2). It may reverse the resistance of tumor cells to conventional chemo- and radiotherapy. Recent research has renewed interest in LDH as an anticancer drug target (3). LDH enzymes have three homologous subunits LDHA, LDHB and LDHC. The combination of these three results to multiple isozymes, including LDH-M and LDH-H. Mutations in LDHB cause lactate dehydrogenase B deficiency, a condition with no deleterious effects on health (4).

References

  1. Faloppi L. et al. (2016) Sci Rep. 6:24136.
  2. Ghosh, M. et al. (2016) Chem. Commun. 52:2401.
  3. Augoff, K. et al. (2015) Cancer Lett. 358:1.
  4. Maekawa M. et al. (1993). Hum. Genet. 91:423.
  5. Maekawa M. et al. (1990). Biochem. Biophys. Res. Commun. 168:677.

Alternate Names

Epididymis Secretory Protein Li 281, HEL-S-281, LDH-H, LDHB, LDHBD, Renal Carcinoma Antigen NY-REN-46, TRG-5

Entrez Gene IDs

3945 (Human); 16832 (Mouse); 24534 (Rat)

Gene Symbol

LDHB

UniProt

P07195

Additional Lactate Dehydrogenase B/LDHB Products

Product Documents for Recombinant Human Lactate Dehydrogenase B/LDHB Protein, CF

Certificate of Analysis

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Note: Certificate of Analysis not available for kit components.

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Product Specific Notices for Recombinant Human Lactate Dehydrogenase B/LDHB Protein, CF

For research use only

Citations for Recombinant Human Lactate Dehydrogenase B/LDHB Protein, CF

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Protocols

View specific protocols for Recombinant Human Lactate Dehydrogenase B/LDHB Protein, CF (9205-HB):

Materials
  • Assay Buffer:  25 mM Tris, 100 mM NaCl, pH 7.5
  • Recombinant Human LDHB (rhLDHB) (Catalog # 9205-HB)
  • beta -Nicotinamide adenine dinucleotide, reduced disodium salt hydrate ( beta -NADH) (Sigma, Catalog # N8129), 20 mM stock in 0.1 M Sodium Borate, pH 9.0
  • 100 mM Sodium Pyruvate (Gibco, Catalog # 11360)
  • 96-well Clear Plate  (Catalog # DY990)
  • Plate Reader (Model: SpectraMax Plus by Molecular Devices) or equivalent
  1. Dilute rhLDHB to 0.4 ng/µL in Assay Buffer.
  2. Prepare a substrate mixture containing 1.6 mM beta -NADH and 4 mM sodium pyruvate in Assay Buffer.
  3. In a plate load 50 µL of 0.4 ng/µL rhLDHB, and start the reaction by adding 50 µL of substrate mixture. Include a Substrate Blank containing 50 µL Assay Buffer and 50 µL of substrate mixture. 
  4. Read plate at 340 nm (absorbance) in kinetic mode for 5 minutes.
  5. Calculate specific activity:

     Specific Activity (pmol/min/µg) =

Adjusted Vmax* (OD/min) x well volume (L) x 1012 pmol/mol x (-1)
ext. coeff** (M-1cm-1) x path corr.*** (cm) x amount of enzyme (µg)


     *Adjusted for Substrate Blank
     **Using the extinction coefficient 6220 M-1cm-1
     ***Using the path correction 0.320 cm
     Note: the output of many spectrophotometers is in mOD.

Per Well:

  • rhLDHB: 0.020 µg
  • beta -NADH: 0.8 mM
  • Sodium pyruvate: 2 mM

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